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Cloning and Transformation of EeHKT1;4 Gene from Elytrigia elongata
Lin Meng1, Lin Zhang, Qiang Guo
1Beijing Research and Development Center for Grass and Environment, Beijing Academy of Agriculture and Forestry Sciences, Haidian District, Beijing, 100097, China. menglin9599@sina.com.
Abstract:
The EeHKT1;4 gene was firstly cloned from Elytrigia elongata by RT-PCR technique with 1977 bp full-length cDNA encoding 1722 bp open reading frame (ORF) and 573 amino acids. The PCR fragment of EeHKT1;4 gene was inserted into the binary vector pBI121 and got the resulted expression vector, which named pBI121-35S-EeHKT1;4-Nos. The vector was further transformed into the agrobacterium EHA105, and then EeHKT1;4 gene was transferred into tobacco by the Agrobaterium- mediated genetic transformation method. The results showed that the target gene was inserted into the genomes of tobacco and expressed. Therefore, the transgenic tobacco (T0) plants overexpressing EeHKT1;4 gene were successfully obtained in this study. And EeHKT1;4 reduces Na+ concentration in the leaves of T0 plants, thereby plays a central role in protecting plant leaves from salinity stress.
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